荧光聚合酶链反应–毛细管电泳法与Sanger测序法检测胶质瘤中端粒酶反转录酶基因启动子突变状态对比分析  

Comparative analysis of telomerase reverse transcriptase gene promoter mutation in glioma between fluorescence polymerase chain reactor-capillary electrophoresis and Sanger sequencing

作　　者：熊艳蕾 王雷明[1] 刘莉 王玮[1] 胡泽良 姚盈盈 段焕利 滕梁红[1] Xiong Yanlei;Wang Leiming;Liu Li;Wang Wei;Hu Zeliang;Yao Yingying;Duan Huanli;Teng Lianghong(Department of Pathology,Xuanwu Hospital,Capital Medical University,Beijing 100053,China)

机构地区：[1]首都医科大学宣武医院病理科,北京100053

出　　处：《北京医学》2022年第3期245-248,共4页Beijing Medical Journal

基　　金：北京市医院管理中心临床医学发展专项——“扬帆”计划(ZYLX202113);首都卫生发展科研专项(首发2020-2-2016)。

摘　　要：目的对比分析荧光聚合酶链反应-毛细管电泳(polymerase chain reaction-capillary electrophoresis,PCR-CE)法及Sanger测序法检测胶质瘤中端粒酶反转录酶(telomerase reverse transcriptase,TERT)基因启动子突变的敏感度、特异度及一致性,为临床检测提供方法学依据。方法收集2017年11月至2019年11月首都医科大学宣武医院265例胶质瘤标本及临床病理资料,分别采用Sanger测序法与荧光PCR-CE法检测TERT基因启动子突变。结果TERT基因启动子突变与年龄、组织学分型和WHO分级均显著相关(P<0.05)。Sanger测序法与荧光PCR-CE法的TERT基因启动子突变检出率分别为52.8%(140/265)及51.3%(136/265),敏感度和特异度分别为96.4%和99.2%,符合率为97.7%,具有较好一致性(Kappa=0.955)。Sanger测序法检出TERT C228T突变103例(38.9%),荧光PCR-CE法检出99例(37.4%),敏感度和特异度分别为95.2%和99.4%,符合率为97.7%,具有较好一致性(Kappa=0.952)。Sanger测序法和荧光PCR-CE法均检测出TERT C250T突变37例(14.0%),符合率为100.0%,具有较高一致性(Kappa=1.000)。结论荧光PCR-CE法检测TERT基因启动子突变率与Sanger测序法相当,敏感度、特异度及一致性均较高,且操作相对简便快速。Objective Compare and analyze the sensitivity,specificity and consistency of fluorescence polymerase chain reaction-capillary electrophoresis(PCR-CE)and Sanger sequencing in the detection of telomerase reverse transcriptase(TERT)gene promoter mutation in gliomas,so as to provide methodological basis for clinical detection.Methods A total of265 glioma samples and clinicopathological data were collected from Xuanwu Hospital,Capital Medical University from November 2017 to November 2019.Mutations of TERT gene promoter were detected by Sanger sequencing and fluorescence PCR-CE,respectively.Results TERT gene promoter mutation was significantly correlated with age,histological type and WHO grade(P<0.05).The detection rates of TERT gene promoter mutation by Sanger sequencing and fluorescence PCR–CE were 52.8%(140/265)and 51.3%(136/265),respectively.The sensitivity and specificity were 96.4%and 99.2%,respectively,and the coincidence rate was 97.7%,with good consistency(Kappa=0.955).One hundred and three cases(38.9%)of TERT C228T mutation were detected by Sanger sequencing and 99 cases(37.4%)by fluorescence PCR–CE.The sensitivity and specificity were 95.2%and 99.4%,respectively,and the coincidence rate was 97.7%,with good consistency(Kappa=0.952).Thirty-seven cases(14.0%)of TERT C250T mutation were detected by Sanger sequencing and fluorescence PCR–CE,and the coincidence rate was 100.0%,with high consistency(Kappa=1.000).Conclusions The mutation rate of TERT gene promoter detected by fluorescence PCR-CE is similar to that by Sanger sequencing,with high sensitivity,specificity and consistency,and the operation is relatively simple and rapid.

关 键 词：胶质瘤 端粒酶反转录酶基因启动子突变 荧光聚合酶链反应-毛细管电泳法 Sanger测序法 

分 类 号：R739.41[医药卫生—肿瘤]